Taheri Reyhaneh, Hadipour Elham, Tayarani-Najaran Zahra
Medical Toxicology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Targeted Drug Delivery Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.
Curr Mol Pharmacol. 2021 Oct 25;14(4):627-634. doi: 10.2174/1874467213666201012160401.
Crocin is a known compound with the antioxidant and anti-inflammatory properties which may help to reduce the progression of neurological disorders. In this study, we aimed to investigate the protective effects of crocin on beta-amyloid peptide Aβ and hydrogen peroxide (HO) induced neurotoxicity in PC12 cells.
PC12 cells were pretreated with crocin and donepezil (5 and 10 μM) for 2 h and then treated with Aβ (25 μM) for 24 h. In parallel, after pretreatment with crocin (5 and 10 μM) and donepezil (5 and 10 μM) for 24 h, cells were treated with H2O2 (800 μM) for 4 h. Finally, the cell viability and intracellular reactive oxygen species (ROS) generation were evaluated using Alamar- Blue® and 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA), respectively. The western blot test was done to compare the protein level of phospho SAPK/JNK, SAPK/JNK, PI3 Kinase P85, Phospho-PI3 Kinase P85, caspase-3 and cytochrome c) cyt c).
Crocin and donepezil could significantly decrease the Aβ toxicity and ROS level. While treatment with Aβ increased Cyt c release from mitochondria to cytosol, cleaved form of caspase-3 (17 kDa) and activated form of SAPK/JNK p44/4 decreased the activated form of PI3 Kinase P85 protein, indicating that crocin could significantly block the apoptosis initiated with Aβ.
According to the results, crocin could be a promising candidate for further evaluations against the development of Alzheimer's disease through mitogen-activated protein kinases (MAPK) and the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signaling (PI3 K/AKT) pathways.
藏红花素是一种已知具有抗氧化和抗炎特性的化合物，可能有助于减缓神经疾病的进展。在本研究中，我们旨在探究藏红花素对β-淀粉样肽Aβ和过氧化氢（H₂O₂）诱导的PC12细胞神经毒性的保护作用。
PC12细胞先用藏红花素和多奈哌齐（5和10μM）预处理2小时，然后用Aβ（25μM）处理24小时。同时，在用藏红花素（5和10μM）和多奈哌齐（5和10μM）预处理24小时后，细胞用H₂O₂（800μM）处理4小时。最后，分别使用阿拉玛蓝®和2',7'-二氯二氢荧光素二乙酸酯（DCFH-DA）评估细胞活力和细胞内活性氧（ROS）的产生。进行蛋白质印迹试验以比较磷酸化SAPK/JNK、SAPK/JNK、PI3激酶P85、磷酸化PI3激酶P85、半胱天冬酶-3和细胞色素c（Cyt c）的蛋白质水平。
藏红花素和多奈哌齐可显著降低Aβ毒性和ROS水平。虽然用Aβ处理会增加细胞色素c从线粒体释放到细胞质中，半胱天冬酶-3的裂解形式（17 kDa）和SAPK/JNK p44/4的活化形式，但PI3激酶P85蛋白的活化形式降低，这表明藏红花素可显著阻断由Aβ引发的细胞凋亡。
根据结果，藏红花素可能是通过丝裂原活化蛋白激酶（MAPK）和磷脂酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）信号传导（PI3 K/AKT）途径进一步评估抗阿尔茨海默病发展的有前途的候选物。
